Substituted pyrimidines for treatment of a cough selected from chronic pathological cough, sub-acute pathological cough, acute pathological cough, neuronal hypersensitivity underlying chronic cough, neuronal hypersensitivity underlying sub-acute cough and neuronal hypersensitivity underlying acute cough

ABSTRACT

Methods for treating cough, chronic cough and urges to cough associated with respiratory diseases with a P2X3 and/or a P2X2/3 receptor antagonist, the methods comprising administering to a subject in need thereof an effective amount of a compound of Formula (I): 
                         
or a pharmaceutically acceptable salt thereof, wherein R 1  and R 2  are as defined herein.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/466,713, filed Aug. 22, 2014, now pending, which claims the benefitunder 35 U.S.C. §119(e) of U.S. Provisional Patent Application Ser. No.61/869,174, filed Aug. 23, 2013, the disclosures of each of which areincorporated herein by reference in their entirety.

FIELD OF THE INVENTION

This invention pertains to compounds and methods for treatment ofdiseases associated with P2 X purinergic receptors, and moreparticularly to methods of using P2X3 and/or P2X2/3 antagonists fortreatment of cough, chronic cough and urge to cough in respiratoryconditions and disorders.

BACKGROUND OF THE INVENTION

The respiratory tract, or airways, participates in the vital process ofgas exchange in order to support the demand for oxygen intake and carbondioxide elimination. Vagal autonomic nerves control smooth muscles ofthe tracheobronchial tree, and thus caliber of airways, as well asliberation and movement of secretions (mucus and fluid). Control iscoordinated within brainstem nuclei which regulate voluntary andautonomic outflow, relying on a rich input of vagal sensory signals fromthe airway tissues that in turn convey conscious sensation and triggerautonomic reflexes. Vagal sensory fibers arise mostly from cell bodieswithin jugular and nodose ganglia, and their activity is regulated by arange of chemical substances (Can & Undem (2003) Respirology8(3):291-301). One such substance is ATP, which sensitizes vagalafferents and serves as a convergent mechanosensory airways signal(Weigand, Ford and Undem (2012) J Physiol. 590(16):4109-20).

ATP activates purinoceptors (e.g., P2X3 and P2X2/3), which mediate manyphysiological and pathological roles (See, Burnstock (1993) Drug Dev.Res. 28:195-206). ATP stimulates and sensitizes sensory nerve endingsresulting in intense sensations such as pain, discomfort, urgency, itchand urge and a pronounced increase in sensory nerve discharge, largelyvia P2X3 receptor activation on afferent nerve fibers innervating rodentand human tissues and organs, especially the hollow viscera.

Data suggest that ATP may be released from epithelial and interstitialcells of hollow organs (such as airways, bladder) as a result ofdistention, movement, pressure or inflammation (Burnstock (1999) J.Anatomy 194:335-342; and Ferguson et al. (1997) J. Physiol.505:503-511). ATP thus serves a role in conveying information to sensoryneurons located in epithelial and subepithelial compartments, e.g.,subepithelial lamina propria (Namasivayam, et al. (1999) BJU Intl.84:854-860; Weigand, Ford and Undem (2012) J Physiol. 590(16):4109-20).

Undem and co-workers have reported that P2X3 and P2X2/3 receptors arewidely expressed and modulate function of nodose and jugular afferentfibers in mammalian airways (Weigand, Ford and Undem (2012) J Physiol.590(16):4109-20). Additionally, in a guinea pig model of ATP orhistamine potentiation of citric acid induced cough, P2X subfamilyreceptors were implicated although contribution of P2X3 or P2X2/3receptors was not deduced (Kamei, Takahashi, Yoshikawa, Saitoh (2005)Eur J Pharmacol. 528(158-161); Kamei and Takahashi. (2006) Eur JPharmacol. 547:160-164). Finally, it has been shown in human studiesthat patients with airway disease associated with cough andbreathlessness (such as asthma, COPD or pulmonary fibrosis) have excessATP concentrations in their airway fluids (Esther, Alexis and Picher.(2011) Subcell. Biochem. 55:75-93; Lommatzsch et al. (2010) Am J RespirCrit Care Med. 181(9):928-34), and that the inhalation by asthmaticpatients of nebulized ATP is able to activate airways sensations leadingto urge to cough and precipitating cough itself (Pellegrino et al.(1996) J Appl Physiol. 81(2):964-75.; Basoglu et al. (2005) Chest.128(4):1905-9), although the site of action of this effect of ATP, andreceptor(s) involved have not been elucidated.

There is accordingly a need for methods of treating diseases, conditionsand disorders mediated by P2X3 and/or P2X2/3 receptors, as well as aneed for compounds that act as modulators of P2X receptors, includingantagonists of P2X3 and P2X2/3 receptors. Such diseases and disordersare herein shown to include cough, chronic cough and urge to cough,including cough associated with a respiratory disease or disorder.Chronic cough is distressing and functionally disabling, and no novellicensed treatments for cough have appeared in approximately 50 years.The present invention satisfies these needs as well as others.

SUMMARY OF THE INVENTION

This invention pertains to compounds and methods of treatment ofdiseases driven by or mediated by P2X3 or P2X2/3 receptor activation,and more particularly to methods of using selective P2X3 and/orP2X3-P2X2/3 antagonists for treatment of common signs, symptoms andmorbidity of diseases mediated by P2X3 and/or P2X2/3 receptors.

The invention provides methods for treating cough-impacted respiratorydiseases using a P2X3 and/or a dual P2X3-P2X2/3 receptor antagonist.More specifically, respiratory diseases can include acute or sub-acutecough, urge to cough, and chronic cough. These respiratory diseases canbe largely corrected by antagonism of P2X3-containing receptors (e.g.,P2X3 and P2X2/3). Moreover, the compounds exemplified herein (e.g.,diaminopyrimidine P2X3/P2X2/3 antagonists) are highly effective atattenuating the cough-related symptoms of many respiratory diseasesincluding acute and sub-acute cough, urge to cough, and chronic cough.

Accordingly, in one aspect, the present invention is directed to amethod for treating a subject for cough or urge to cough associated witha respiratory disease. The method can comprise administering to thesubject in need thereof an effective amount of a compound of Formula(I):

or a pharmaceutically acceptable salt thereof.

In one or more embodiments, R¹ is hydrogen or optionally substitutedC₁-C₆ alkyl. In one or more embodiments, R² is: alkyl; alkenyl; alkynyl;amino; aminosulfonyl; halo; amido; haloalkyl; alkoxy; hydroxy;haloalkoxy; nitro; hydroxyalkyl; alkoxyalkyl; hydroxyalkoxy;alkynylalkoxy; alkylsulfonyl; arylsulfonyl; carboxyalkyl; cyano oralkylcarbonyl.

In one or more embodiments, the respiratory symptom, condition ordisorder is attenuated by a P2X3 or P2X3-P2X2/3 receptor antagonist. Therespiratory disease can be selected from many conditions where coughhypersensitivity prevails, and may include unexplained cough or coughassociated with upper respiratory infection, chronic obstructivepulmonary disease (COPD), asthma, idiopathic pulmonary fibrosis.

In one or more embodiments, the cough is sub-acute or chronic cough,treatment-resistant cough, idiopathic chronic cough, post-viral cough,iatrogenic cough, cough associated with post-nasal drip, coughassociated with upper respiratory infection, asthma and/or COPD, coughassociated with interstitial disease, cough associated withgastroesophageal reflux disease (GERD) and/or cough associated withsmoking or a form of bronchitis. The iatrogenic cough can be induced byan ACE-inhibitor. Additionally, the interstitial disease can bepulmonary fibrosis.

In another aspect, the present invention is directed to a method fortreating chronic cough in a patient in need thereof. The method cancomprise administering an effective amount of a compound of Formula (I):

or a pharmaceutically acceptable salt thereof. In one or moreembodiments, R¹ is hydrogen or optionally substituted C₁-C₆ alkyl.

In one or more embodiments, R² is: alkyl; alkenyl; alkynyl; amino;aminosulfonyl; halo; amido; haloalkyl; alkoxy; hydroxy; haloalkoxy;nitro; hydroxyalkyl; alkoxyalkyl; hydroxyalkoxy; alkynylalkoxy;alkylsulfonyl; arylsulfonyl; carboxyalkyl; cyano or alkylcarbonyl.

In one or more embodiments, the chronic cough is idiopathic or treatmentresistant cough. The cough can be daytime cough.

In another aspect, the present invention provides a method for treatingneuronal hypersensitivity underlying acute, sub-acute or chronic cough.The method comprises administering an effective amount of a compound ofFormula (I):

or a pharmaceutically acceptable salt thereof.

In one or more embodiments, R¹ is hydrogen or optionally substitutedC₁-C₆ alkyl.

In one or more embodiments, R² is: alkyl; alkenyl; alkynyl; amino;aminosulfonyl; halo; amido; haloalkyl; alkoxy; hydroxy; haloalkoxy;nitro; hydroxyalkyl; alkoxyalkyl; hydroxyalkoxy; alkynylalkoxy;alkylsulfonyl; arylsulfonyl; carboxyalkyl; cyano or alkylcarbonyl.

In one or more embodiments, the chronic cough is idiopathic or treatmentresistant cough. The cough can be daytime cough.

In some embodiments of any of the above aspects, R¹ can be methyl orhydrogen. In some embodiments of any of the above aspects, R² can behaloalkyl, aminosulfonyl, alkylsulfonyl alkylcarbonyl or carboxyalkyl.In some embodiments, R² is haloalkyl, further wherein the alkyl ismethyl. R² can also be aminosulfonyl, carboxyalkyl, or alkylcarbonyl.

In one or more embodiments, of any of the above aspects, the compound ofFormula (I) is administered at about 600 mg twice daily. The compound ofFormula (I) can be administered for about 2 weeks. For instance, thecompound of Formula (I) is administered at about 600 mg twice daily forabout two weeks.

In one or more embodiments of any of the above aspects, the chroniccough is refractory chronic cough. In one or more embodiments, of any ofthe above aspects, the chronic cough is reduced by about 50-90% (e.g.,about 55%, 60%, 65%, 70%, 75%, 80%, or 85%).

In one or more embodiments of any of the above aspects, the P2X3 orP2X2/3 antagonist compound is selected from Compounds 1-38. Forinstance, the compound can be selected from Compounds 6, 7, 13, 16, 20,27, 34 and 37 (e.g., the compound can be Compound 16).

In one or more embodiments, the invention relates to a method fortreating the symptoms of cough and urge to cough associated with arespiratory disease by administering a compound of Formula (I). Forexample, the invention relates to a method of treatment of the symptomsof chronic cough and/or urge to cough associated with a respiratorydisease or disorder mediated by a P2X3 or P2X2/3 receptor antagonist byadministering a compound of Formula (I).

In one or more embodiments, the invention relates to methods forreducing daytime chronic cough in idiopathic/treatment-resistant chroniccough. The invention also relates to a method of treating neuronalhypersensitivity underlying chronic cough.

In one or more embodiments, the methods of the invention relate totreating, preventing or ameliorating the respiratory diseases anddisorders described herein, or symptoms thereof, described herein in apatient in need thereof by administering a compound selected fromCompounds 1-39. For example, the compound is selected from Compounds 6,7, 13, 17, 21, 28, 35 and 38. For example, the compound is Compound 16.

The invention also provides pharmaceutical compositions of the compoundsof the present invention and methods of preparing the same.

As set forth in the Detailed Description below, the present inventionfeatures a class of P2X3 and P2X2/3 antagonists for treating oralleviating cough and urge to cough, including chronic cough. Thepresent invention has the advantage of addressing the root cause drivingcough hypersensitivity in these illnesses instead of merely suppressingcentral modulation of the symptom perception. For instance, the presentinvention offers methods to reduce the activity of afferent nerves thatultimately trigger the persistent and inappropriate urge to cough in asensitized subject (e.g., a human). The present invention also has theadvantage of giving highly selective P2X3 and P2X2/3 antagonists.Further features and advantages are set forth in the DetailedDescription below and will be apparent to one of skill in the art.

BRIEF DESCRIPTION OF THE DRAWING

The FIGURE shows a graph depicting the effect of Compound 16 onobjectively recorded daytime cough frequency (coughs per hour) intreatment-resistant chronic cough patients.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

Unless otherwise stated, the following terms used in this Application,including the specification and claims, have the definitions givenbelow. It must be noted that, as used in the specification and theappended claims, the singular forms “a”, “an,” and “the” include pluralreferents unless the context clearly dictates otherwise.

“Agonist” refers to a compound that enhances the activity of anothercompound or receptor site.

“Alkyl” means the monovalent linear or branched saturated hydrocarbonmoiety, consisting solely of carbon and hydrogen atoms, having from oneto twelve carbon atoms. “Lower alkyl” refers to an alkyl group of one tosix carbon atoms, i.e. C₁-C₆alkyl. Examples of alkyl groups include, butare not limited to, methyl, ethyl, propyl, isopropyl, isobutyl,sec-butyl, tert-butyl, pentyl, n-hexyl, octyl, dodecyl, and the like.

“Alkenyl” means a linear monovalent hydrocarbon radical of two to sixcarbon atoms or a branched monovalent hydrocarbon radical of three tosix carbon atoms, containing at least one double bond, e.g., ethenyl,propenyl, and the like.

“Alkynyl” means a linear monovalent hydrocarbon radical of two to sixcarbon atoms or a branched monovalent hydrocarbon radical of three tosix carbon atoms, containing at least one triple bond, e.g., ethynyl,propynyl, and the like.

“Alkylene” means a linear saturated divalent hydrocarbon radical of oneto six carbon atoms or a branched saturated divalent hydrocarbon radicalof three to six carbon atoms, e.g., methylene, ethylene,2,2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene,and the like.

“Alkoxy” means a moiety of the formula —OR, wherein R is an alkyl moietyas defined herein. Examples of alkoxy moieties include, but are notlimited to, methoxy, ethoxy, isopropoxy, and the like.

“Alkoxyalkyl” means a moiety of the formula R^(a)—O—R^(b)—, where R^(a)is alkyl and R^(b) is alkylene as defined herein. Exemplary alkoxyalkylgroups include, by way of example, 2-methoxyethyl, 3-methoxypropyl,1-methyl-2-methoxyethyl, 1-(2-methoxyethyl)-3-methoxypropyl, and1-(2-methoxyethyl)-3-methoxypropyl.

“Alkylcarbonyl” means a moiety of the formula —R′—R″, where R′ is (C═O)and R″ is alkyl as defined herein.

“Alkylsulfonyl” means a moiety of the formula —R′—R″, where R′ is —SO₂—and R″ is alkyl as defined herein.

“Alkylsulfonylalkyl” means a moiety of the formula —R′—R″—R′″ where R′is alkylene, R″ is —SO₂— and R′″ is alkyl as defined herein.

“Alkylamino” means a moiety of the formula —NR—R′ wherein R is hydrogenor alkyl and R′ is alkyl as defined herein.

“Alkylsulfanyl” means a moiety of the formula —SR wherein R is alkyl asdefined herein.

“Amino” means a moiety of the formula —NHR wherein R can be hydrogen oralkyl.

“Amido” means a moiety of the formula —NR(CO)R′— wherein R and R′ can beH or alkyl as defined herein.

“Hydroxy” means a moiety of the formula —OH.

“Haloalkoxy” means a group of the formula —OR, wherein R is a haloalkylgroup as defined herein.

“Nitro” means a group of the formula —NO₂. “Alkylcarbonyl” refers to agroup of the formula —(CO)R wherein R is an alkyl group as definedherein.

“Aminoalkyl” means a group—R-R′ wherein R′ is amino and R is alkylene asdefined herein. “Aminoalkyl” includes aminomethyl, aminoethyl,1-aminopropyl, 2-aminopropyl, and the like. The amino moiety of“aminoalkyl” may be substituted once or twice with alkyl to provide“alkylaminoalkyl” and “dialkylaminoalkyl” respectively.“Alkylaminoalkyl” includes methylaminomethyl, methylaminoethyl,methylaminopropyl, ethylaminoethyl and the like. “Dialkylaminoalkyl”includes dimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl,N-methyl-N-ethylaminoethyl, and the like.

“Aminosulfonyl” means a group —SO₂—NRR′ wherein R and R′ eachindependently is hydrogen or alkyl as defined herein.

“Alkylsulfonylamido” means a moiety of the formula —NR'SO₂—R wherein Ris alkyl and R′ is hydrogen or alkyl.

“Alkynylalkoxy” means a group of the formula —O—R—R′ wherein R isalkylene and R′ is alkynyl as defined herein.

“Aryl” means a monovalent cyclic aromatic hydrocarbon moiety consistingof a mono-, bi- or tricyclic aromatic ring. The aryl group can beoptionally substituted as defined herein. Examples of aryl moietiesinclude, but are not limited to, optionally substituted phenyl,naphthyl, phenanthryl, fluorenyl, indenyl, pentalenyl, azulenyl,oxydiphenyl, biphenyl, methylenediphenyl, aminodiphenyl,diphenylsulfidyl, diphenylsulfonyl, diphenylisopropylidenyl,benzodioxanyl, benzofuranyl, benzodioxylyl, benzopyranyl, benzoxazinyl,benzoxazinonyl, benzopiperadinyl, benzopiperazinyl, benzopyrrolidinyl,benzomorpholinyl, methylenedioxyphenyl, ethylenedioxyphenyl, and thelike, including partially hydrogenated derivatives thereof.

“Arylsulfonyl” means a group of the formula —SO₂—R wherein R is aryl asdefined herein.

“Antagonist” refers to a compound that diminishes or prevents the actionof another compound or receptor site.

“Cyanoalkyl” “means a moiety of the formula —R′—R”, where R′ is alkyleneas defined herein and R″ is cyano or nitrile.

The terms “halo”, “halogen” and “halide”, which may be usedinterchangeably, refer to a substituent fluoro, chloro, bromo, or iodo.

“Haloalkyl” means alkyl as defined herein in which one or more hydrogenatoms have been replaced with the same or different halogen. Exemplaryhaloalkyls include —CH₂Cl, —CH₂CF₃, —CH₂CCl₃, perfluoroalkyl (e.g.,—CF₃), and the like.

“Hydroxyalkoxy” means a moiety of the formula —OR wherein R ishydroxyalkyl as defined herein.

“Hydroxyalkylamino” means a moiety of the formula —NR—R′ wherein R ishydrogen or alkyl and R′ is hydroxyalkyl as defined herein.

“Hydroxyalkylaminoalkyl” means a moiety of the formula —R—NR′—R″ whereinR is alkylene, R′ is hydrogen or alkyl, and R″ is hydroxyalkyl asdefined herein.

“Hydroxycarbonylalkyl” or “carboxyalkyl” means a group of the formula—R—(CO)—OH where R is alkylene as defined herein.

“Hydroxyalkyloxycarbonylalkyl” or “hydroxyalkoxycarbonylalkyl” means agroup of the formula —R—C(O)—O—R—OH wherein each R is alkylene and maybe the same or different.

“Hydroxyalkyl” means an alkyl moiety as defined herein, substituted withone or more, preferably one, two or three hydroxy groups, provided thatthe same carbon atom does not carry more than one hydroxy group.Representative examples include, but are not limited to, hydroxymethyl,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl,4-hydroxybutyl, 2,3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl,2,3-dihydroxybutyl, 3,4-dihydroxybutyl and2-(hydroxymethyl)-3-hydroxypropyl

“Carboxy” means a group of the formula —O—C(O)—OH.

“Sulfonamido” means a group of the formula —SO₂—NR′R″ wherein R′ and R″each independently is hydrogen or alkyl.

“Optionally substituted”, for example when used with the term alkyl,means an alkyl group which is optionally substituted independently withone to three substituents, preferably one or two substituents selectedfrom any of the substituents defined herein, for instance alkyl,cycloalkyl, cycloalkylalkyl, heteroalkyl, hydroxyalkyl, halo, nitro,cyano, hydroxy, alkoxy, amino, acylamino, mono-alkylamino,di-alkylamino, haloalkyl, haloalkoxy, heteroalkyl, —COR (where R ishydrogen, alkyl, phenyl or phenylalkyl), —(CR′R″)_(n)—COOR (where n isan integer from 0 to 5, R′ and R″ are independently hydrogen or alkyl,and R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl orphenylalkyl), or —(CR′R″)_(n)—CONR^(a)R^(b) (where n is an integer from0 to 5, R′ and R″ are independently hydrogen or alkyl, and R^(a) andR^(b) are, independently of each other, hydrogen, alkyl, cycloalkyl,cycloalkylalkyl, phenyl or phenylalkyl).

“Leaving group” means the group with the meaning conventionallyassociated with it in synthetic organic chemistry, i.e., an atom orgroup displaceable under substitution reaction conditions. Examples ofleaving groups include, but are not limited to, halogen, alkane- orarylenesulfonyloxy, such as methanesulfonyloxy, ethanesulfonyloxy,thiomethyl, benzenesulfonyloxy, tosyloxy, and thienyloxy,dihalophosphinoyloxy, optionally substituted benzyloxy, isopropyloxy,acyloxy, and the like.

“Modulator” means a molecule that interacts with a target. Theinteractions include, but are not limited to, agonist, antagonist, andthe like, as defined herein.

“Optional” or “optionally” means that the subsequently described eventor circumstance may but need not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not.

“Disease” and “Disease state” means any disease, condition, symptom,disorder or indication.

“Inert organic solvent” or “inert solvent” means the solvent is inertunder the conditions of the reaction being described in conjunctiontherewith, including for example, benzene, toluene, acetonitrile,tetrahydrofuran, N,N-dimethylformamide, chloroform, methylene chlorideor dichloromethane, dichloroethane, diethyl ether, ethyl acetate,acetone, methyl ethyl ketone, methanol, ethanol, propanol, isopropanol,tert-butanol, dioxane, pyridine, and the like. Unless specified to thecontrary, the solvents used in the reactions of the present inventionare inert solvents.

“Pharmaceutically acceptable” means that which is useful in preparing apharmaceutical composition that is generally safe, non-toxic, andneither biologically nor otherwise undesirable and includes that whichis acceptable for veterinary as well as human pharmaceutical use.

“Pharmaceutically acceptable salts” of a compound means salts that arepharmaceutically acceptable, as defined herein, and that possess thedesired pharmacological activity of the parent compound. Such saltsinclude: acid addition salts formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like; or formed with organic acids such asacetic acid, benzenesulfonic acid, benzoic, camphorsulfonic acid, citricacid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconicacid, glutamic acid, glycolic acid, hydroxynaphtoic acid,2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid,malonic acid, mandelic acid, methanesulfonic acid, muconic acid,2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinicacid, tartaric acid, p-toluenesulfonic acid, trimethylacetic acid, andthe like; or salts formed when an acidic proton present in the parentcompound either is replaced by a metal ion, e.g., an alkali metal ion,an alkaline earth ion, or an aluminum ion; or coordinates with anorganic or inorganic base. Acceptable organic bases includediethanolamine, ethanolamine, N-methylglucamine, triethanolamine,tromethamine, and the like. Acceptable inorganic bases include aluminumhydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate andsodium hydroxide.

The preferred pharmaceutically acceptable salts are the salts formedfrom acetic acid, hydrochloric acid, sulphuric acid, methanesulfonicacid, maleic acid, phosphoric acid, tartaric acid, citric acid, sodium,potassium, calcium, zinc, and magnesium.

It should be understood that all references to pharmaceuticallyacceptable salts include solvent addition forms (solvates) or crystalforms (polymorphs) as defined herein, of the same acid addition salt.

“Protective group” or “protecting group” means the group whichselectively blocks one reactive site in a multifunctional compound suchthat a chemical reaction can be carried out selectively at anotherunprotected reactive site in the meaning conventionally associated withit in synthetic chemistry. Certain processes of this invention rely uponthe protective groups to block reactive nitrogen and/or oxygen atomspresent in the reactants. For example, the terms “amino-protectinggroup” and “nitrogen protecting group” are used interchangeably hereinand refer to those organic groups intended to protect the nitrogen atomagainst undesirable reactions during synthetic procedures. Exemplarynitrogen protecting groups include, but are not limited to,trifluoroacetyl, acetamido, benzyl (Bn), benzyloxycarbonyl(carbobenzyloxy, CBZ), p-methoxybenzyloxycarbonyl,p-nitrobenzyloxycarbonyl, tert-butoxycarbonyl (BOC), and the like. Theartisan in the art will know how to choose a group for the ease ofremoval and for the ability to withstand the following reactions.

“Solvate” or “solvates” means solvent addition forms that contain eitherstoichiometric or non-stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate; when the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one of the substances in whichthe water retains its molecular state as H₂O, such combination beingable to form one or more hydrate.

“Subject” means mammals and non-mammals. Mammals means any member of themammalia class including, but not limited to, humans; non-human primatessuch as chimpanzees and other apes and monkey species; farm animals suchas cattle, horses, sheep, goats, and swine; domestic animals such asrabbits, dogs, and cats; laboratory animals including rodents, such asrats, mice, and guinea pigs; and the like. Examples of non-mammalsinclude, but are not limited to, birds, and the like. The term “subject”does not denote a particular age or sex.

“Cough related respiratory disorder” or “respiratory disease” refers to,without limitation, cough hypersensitivity syndrome, chronic obstructivepulmonary disease (COPD), asthma, bronchospasm, and the like.Respiratory disorders include, for example, sub-acute or chronic cough,treatment-resistant cough, idiopathic chronic cough, cough associatedwith upper respiratory infection, post-viral cough, iatrogenic cough(e.g., as induced by ACE-inhibitors), idiopathic pulmonary fibrosis orcough associated with smoking or a form of bronchitis. Respiratorydisorders can include urge to cough associated with any respiratorydisease, for example urge to cough associated with chronic obstructivepulmonary disease (COPD), cough-variant asthma, interstitial lungdisease, or whooping cough.

“Acute cough” is understood to mean a cough lasting up to two weeks induration. For instance, acute cough can be the result of an acutedisease, such as a cold or flu. An acute cough will disappear when theunderlying cause (e.g., cold or flu) is eliminated.

“Sub-acute cough” is understood to mean a cough lasting between two andeight weeks. In some cases, a sub-acute cough follows a period in whicha subject is infected with a disease (e.g., cold or flu). A sub-acutecough is one that often remains after the underlying cause has beenremoved. For instance, a sub-acute cough is found post-infection (e.g.,post-viral infection).

“Chronic cough” refers to a persistent or refractory cough lastinglonger than eight weeks that may not have an obvious underlying causeand is may not be associated with other respiratory diseases, such asasthma or COPD (i.e., idiopathic). Chronic cough is also characterizedin that there are no hallmarks to define and diagnose it, in contrast toother respiratory diseases (e.g., COPD). Another characteristic ofchronic cough is that a subject suffering from chronic cough may beapparently normal in most other respects. Chronic cough is characterizedby frequent coughing (e.g., at least 5-10 coughs per hour duringdaytime), and bothersome coughing during sleep. Chronic cough can lastfor a period of years, including over a decade.

In order to determine if a subject is afflicted by a chronic cough, apractitioner or clinician can perform a three-step test. First, thesubject can be treated for putative post-nasal drips. In some cases,such treatment takes the form of an antihistamine. Second, the subjectcan be treated with a proton-pump inhibitor (e.g., to treat putativegastro-esophageal disease such as reflux disease). Third, a subject canbe treated with steroids (e.g., to treat a putative case of asthma).

If a subject continues to display a chronic cough after the abovethree-step treatment regimen, the cough is said to be chronic cough andis likely refractory. It is understood that patients suffering fromrefractory cough often have suffered both acute and sub-acute coughbefore being diagnosed with chronic cough.

“Therapeutically effective amount” means an amount of a compound that,when administered to a subject for treating a disease state, issufficient to effect such treatment for the disease state. The“therapeutically effective amount” will vary depending on the compound,disease state being treated, the severity or the disease treated, theage and relative health of the subject, the route and form ofadministration, the judgment of the attending medical or veterinarypractitioner, and other factors.

The terms “those defined above” and “those defined herein” whenreferring to a variable incorporates by reference the broad definitionof the variable as well as preferred, more preferred and most preferreddefinitions, if any.

“Treating” or “treatment” of a disease state includes:

-   -   (i) inhibiting the disease state, i.e., arresting the        development of the disease state or its clinical symptoms, or    -   (ii) relieving the disease state, i.e., causing temporary or        permanent regression of the disease state or its clinical        symptoms.

“Preventing” or “prevention” of a disease state includes causing theclinical symptoms of the disease state not to develop in a subject thatmay be exposed to or predisposed to the disease state, but does not yetexperience or display symptoms of the disease state. For example,treating or preventing a respiratory disease or disorder includestreating or preventing the symptoms the disorder such as cough and/orurge to cough associated with a respiratory disease.

The terms “treating”, “contacting” and “reacting” when referring to achemical reaction means adding or mixing two or more reagents underappropriate conditions to produce the indicated and/or the desiredproduct. It should be appreciated that the reaction which produces theindicated and/or the desired product may not necessarily result directlyfrom the combination of two reagents which were initially added, i.e.,there may be one or more intermediates which are produced in the mixturewhich ultimately leads to the formation of the indicated and/or thedesired product.

Nomenclature and Structures

In general, the nomenclature used in this Application is based onAUTONOM™ v.4.0; a Beilstein Institute computerized system for thegeneration of IUPAC systematic nomenclature. Chemical structures shownherein were prepared using ISIS® version 2.2. Any open valency appearingon a carbon, oxygen or nitrogen atom in the structures herein indicatesthe presence of a hydrogen atom.

All patents and publications identified herein are incorporated hereinby reference in their entirety.

Methods

The invention provides compounds and methods for treating a respiratorydisease mediated by a P2X3 or P2X2/3 receptor antagonist, said methodcomprising administering to a subject in need thereof an effectiveamount of a compound of Formula (I):

wherein:

R¹ is hydrogen or optionally substituted C₁-C₆ alkyl;

R² is: alkyl; alkenyl; alkynyl; amino; aminosulfonyl; halo; amido;haloalkyl; alkoxy; hydroxy; haloalkoxy; nitro; hydroxyalkyl;alkoxyalkyl; hydroxyalkoxy; alkynylalkoxy; alkylsulfonyl; arylsulfonyl;carboxyalkyl; cyano or alkylcarbonyl.

Exemplary respiratory diseases treatable with the compounds and methodsof the invention include acute, sub-acute or chronic cough,treatment-resistant cough, idiopathic chronic cough, post-viral cough,whooping couh, iatrogenic cough (e.g., as induced by ACE-inhibitors),idiopathic pulmonary fibrosis or cough associated with smoking or a formof bronchitis. A disease treatable by the invention includes urge tocough associated with any respiratory disease, for example urge to coughassociated with chronic obstructive pulmonary disease (COPD), or asthma.For example, the invention relates to a method for treating the symptomsof cough and urge to cough associated with a respiratory disease. Forexample, the invention relates to a method of treatment of the symptomsof cough and/or urge to cough associated with a respiratory disease ordisorder mediated by a P2X3 or P2X2/3 receptor antagonist.

The present invention also provides methods of treatment of arespiratory disease, wherein the respiratory symptoms are mediated byP2X3 and/or P2X2/3 receptor activation. The method can includeadministering to the subject an effective amount of a compound ofFormula (I). The method can include administering any of the embodimentsof Formula (I) set forth herein.

The present invention also provides compounds for use in treatingrespiratory symptoms mediated by P2X3 and/or P2X2/3 receptor activation(e.g., cough or chronic cough). The present invention also provides thatcompounds of the present invention can be used in the manufacture of amedicament for use in treating a respiratory disease mediated by P2X3and/or P2X2/3 receptor activation and senstization (e.g., cough orchronic cough).

In certain embodiments of the invention the respiratory disease to betreated or prevented may be chronic cough. For example, the inventionrelates to methods for reducing daytime cough inidiopathic/treatment-resistant chronic cough. In some embodiments, asubject with chronic cough has as many as 40 coughs per hour or moreover a period of 24 hours (e.g., at least 25 coughs per hour). In someembodiments, the chronic cough is not obviously caused by an underlyingdisease or ailment. For instance, the chronic cough can be caused bypersistent endogenous over-activation of a P2X3 or a P2X2/3 receptor.Such activation may not be the result of a separate ailment. In certainembodiments of the invention, the symptom or disorder to be treated isneuronal hypersensitivity underlying chronic cough.

Without wishing to be bound by any particular theory, in someembodiments, the present invention can work by antagonizing andultimately modulating (e.g., reducing) the activity of P2X3 and/orP2X2/3 receptors. This in turn can have the effect of modulating (e.g.,down regulating) the function of nodose and jugular afferent fibers inmammalian airways. This process can have the global effect of reducingthe neuronal signals that trigger the urge to cough, e.g., in a patientsuffering from acute, sub-acute or chronic cough. Without wishing to bebound by any theory, Compound 16 may be an efficacious treatment forchronic coughing. Further without wishing to be bound by theory, theP2X3 receptor modulators described herein can adjust and/or attenuatethe neuronal hypersensitivity underlying chronic cough.

In many embodiments of the invention the disorder to be treated orprevented is urge to cough associated with a respiratory disease.

For example, the methods of the invention relate to treating, preventingor ameliorating the respiratory diseases and disorders described herein,or symptoms thereof, described herein in a patient in need thereof byadministering a compound selected from Compounds 1-39. For example, thecompound is selected from Compounds 6, 7, 13, 17, 21, 28, 35 and 38. Forexample, the compound is Compound 16.

In some instances, preferred embodiments from one group can be combinedwith preferred embodiments from another group. For instance, in onepreferred embodiment R₁ is —CH₃. In another preferred embodiment, R₂ is—SO₂NH₂. According to the present disclosure, the two preferredembodiments disclosed above can be combined to give a preferred compoundwherein R₁ is —CH₃ and R₂ is —SO₂NH₂.

In certain embodiments of Formula (I), R¹ is methyl.

In certain embodiments of Formula (I), R¹ is hydrogen.

In certain embodiments of Formula (I), R² is haloalkyl, aminosulfonyl,alkylsulfonyl alkylcarbonyl or carboxyalkyl.

In certain embodiments of Formula (I), R² is haloalkyl, where alkyl ismethyl.

In certain embodiments of Formula (I), R² is aminosulfonyl.

In certain embodiments of Formula (I), R² is carboxyalkyl.

In certain embodiments of Formula (I), R² is alkylcarbonyl.

Where R¹ or R² is alkyl or contains an alkyl moiety, such alkyl ispreferably lower alkyl, i.e. C₁-C₆alkyl, and more preferably C₁-C₄alkyl.

Representative compounds in accordance with the methods of the inventionare shown in Table 1.

TABLE 1 # Structure Name  1

5-(2-Isopropyl-4,5-dimethoxy-phenoxy)- pyrimidine-2,4-diamine  2

5-(5-Bromo-2-isopropyl-4-methoxy- phenoxy)-pyrimidine-2,4-diamine  3

5-(5-Chloro-2-isopropyl-4-methoxy- phenoxy)-pyrimidine-2,4-diamine  4

5-(2-Isopropyl-4-methoxy-5-methyl- phenoxy)-pyrimidine-2,4-diamine  5

1-[5-(2,4-Diamino-pyrimidin-5-yloxy)- 4-isopropyl-2-methoxy-phenyl]-ethanone  6

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-benzamide  7

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-benzoic acid  8

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-benzonitrile  9

[5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-phenyl]-urea10

5-(5-Chloro-4-difluoromethoxy-2- isopropyl-phenoxy)-pyrimidine-2,4-diamine 11

5-(5-Amino-2-isopropyl-4-methoxy- phenoxy)-pyrimidine-2,4-diamine 12

N-[5-(2,4-Diamino-pyrimidin-5-yloxy)- 4-isopropyl-2-methoxy-phenyl]-acetamide 13

5-(2-Isopropyl-5-methanesulfonyl-4- methoxy-phenoxy)-pyrimidine-2,4-diamine 14

1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-2-hydroxy-4-isopropyl-phenyl]-ethanone 15

5-(5-Iodo-2-isopropyl-4-methoxy- phenoxy)-pyrimidine-2,4-diamine 16

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-benzenesulfonamide 17

4-(2,4-Diamino-pyrimidin-5-yloxy)-2- iodo-5-isopropyl-phenol 18

5-(2-Isopropyl-4methoxy-5-vinyl- phenoxy)-pyrimidine-2,4-diamine 19

4-(2,4-Diamino-pyrimidin-5-ylmethyl)- 2-iodo-5-isopropyl-phenol 20

5-(2-Isopropyl-4-methoxy-5- trifluoromethyl-phenoxy)-pyrimidine-2,4-diamine 21

1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-3-ethyl- urea 22

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-N-methyl-benzamide 23

1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanol 24

5-(2,5-Diisopropyl-4-methoxy- phenoxy)-pyrimidine-2,4-diamine 25

5-[2-Isopropyl-4-methoxy-5-(1- methoxy-ethyl)-phenoxyl-pyrimidine-2,4-diamine 26

1-[5-(2,4-Diamino-pyrimidin-5-yloxy)- 4-isopropyl-2-methoxy-phenyl]-3-phenyl-urea 27

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-N-methyl-benzenesulfonamide 28

5-(2-Isopropyl-4-methoxy-5- trifluoromethoxy-phenoxy)-pyrimidine-2,4-diamine 29

5-(5-Iodo-2-isopropyl-4-prop-2- ynyloxy-phenoxy)-pyrimidine-2,4- diamine30

5-(2-Isopropyl-4-methoxy-5-nitro- phenoxy)-pyrimidine-2,4-diamine 31

5-(4-Ethoxy-5-iodo-2-isopropyl- phenoxy)-pyrimidine-2,4-diamine 32

5-[5-Iodo-2-isopropyl-4-(2,2,2-trifluoro-ethoxy)-phenoxy]-pyrimidine-2,4- diamine 33

5-(2-Isopropyl-4-methoxy-5-nitro- phenoxy)-pyrimidine-2,4-diamine 34

5-(5-Ethanesulfonyl-2-isopropyl-4- methoxy-phenoxy)-pyrimidine-2,4-diamine 35

5-(5-Fluoro-2-isopropyl-4-methoxy- phenoxy)-pyrimidine-2,4-diamine 36

2-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-propan- 2-ol 37

5-(2,4-Diamino-pyrimidin-5-yloxy)-N- ethyl-4-isopropyl-2-methoxy-benzenesulfonamide 38

5-(2,4-Diamino-pyrimidin-5-yloxy)-4- isopropyl-2-methoxy-N,N-dimethyl-benzamideSynthesis

Compounds of the present invention can be made by a variety of methodsdepicted in the illustrative synthetic reaction schemes shown anddescribed below. Syntheses of compounds for use in the invention canalso be performed according to teachings presented in, for example, U.S.Pat. Nos. 7,858,632, 8,008,313; 8,003,788; 7,531,547; 7,741,484 and7,799,796, each of which is specifically incorporated herein in itsentirety.

The starting materials and reagents used in preparing these compoundsgenerally are either available from commercial suppliers, such asAldrich Chemical Co., or are prepared by methods known to those skilledin the art following procedures set forth in references such as Fieserand Fieser's Reagents for Organic Synthesis; Wiley & Sons: New York,1991, Volumes 1-15; Rodd's Chemistry of Carbon Compounds, ElsevierScience Publishers, 1989, Volumes 1-5 and Supplementals; and OrganicReactions, Wiley & Sons: New York, 1991, Volumes 1-40. The followingsynthetic reaction schemes are merely illustrative of some methods bywhich the compounds of the present invention can be synthesized, andvarious modifications to these synthetic reaction schemes can be madeand will be suggested to one skilled in the art having referred to thedisclosure contained in this Application.

The starting materials and the intermediates of the synthetic reactionschemes can be isolated and purified if desired using conventionaltechniques, including but not limited to, filtration, distillation,crystallization, chromatography, and the like. Such materials can becharacterized using conventional means, including physical constants andspectral data.

Unless specified to the contrary, the reactions described hereinpreferably are conducted under an inert atmosphere at atmosphericpressure at a reaction temperature range of from about −78° C. to about150° C., more preferably from about 0° C. to about 125° C., and mostpreferably and conveniently at about room (or ambient) temperature,e.g., about 20° C.

Scheme A below illustrates another synthetic procedure usable to preparespecific compounds of Formula (I) above, wherein R³, R⁴, R^(d), andR^(e) are as defined herein.

In step 1 of Scheme A, an O-alkylation is carried out by reaction ofphenol i with a haloacetonitrile such as iodoacetonitrile k, to affordcyano ether l. Numerous substituted phenols i are either commerciallyavailable or may be prepared by techniques well known in the art for usein step 1. For example, substituted aldehydes may be converted to thecorresponding phenols i via Baeyer-Villiger oxidation using peracid suchas mCPBA, as illustrated in the experimental examples below. Thealkylation of step 1 may be effected in the presence of mild base underpolar aprotic solvent conditions.

In step 2, a cyano enol ether compound n is formed by treatment of cyanoether l with a strong base such as sodium hydride, followed byintroduction of ester m to form an enolate (not shown), that in turn isalkylated by addition of iodomethane or other alkyl halide. This stepmay be carried out under polar aprotic solvent conditions.

In step 3 cyano enol ether n is reacted with guanidine compound o in thepresence of base, under polar aprotic conditions, to yielddiaminopyrimidine (p). The diaminopyrimidine (p) is a compound ofFormula (I) usable in the methods of the invention.

Numerous variations on the procedure of Scheme A are possible and willbe readily apparent to those skilled in the art.

Specific details for producing compounds of the invention are describedin the Examples section below.

Use

The invention provides methods for treating a respiratory diseasemediated by a P2X₃ or P2X_(2/3) receptor antagonist, said methodcomprising administering to a subject in need thereof an effectiveamount of a compound of Formula (I):

Exemplary respiratory diseases treatable with the invention includesub-acute or chronic cough, treatment-resistant cough, idiopathicchronic cough, post-viral cough, iatrogenic cough (e.g., as induced byACE-inhibitors), idiopathic pulmonary fibrosis or cough associated withsmoking or a form of bronchitis. A disease treatable by the inventionincludes urge to cough associated with any respiratory disease, forexample urge to cough associated with chronic obstructive pulmonarydisease (COPD), asthma or bronchospasm.

In certain embodiments of the invention the respiratory disease may bechronic cough.

In many embodiments of the invention the disease is urge to coughassociated with a respiratory disease.

In one or more embodiments, the compounds of the present invention canresult in a reduction of objective daytime cough counts in a subjectwith chronic cough (e.g., refractory chronic cough). For instance,administration of the compounds of the present invention or apharmaceutically acceptable salt thereof to a subject in need thereofcan result in a reduction in objective daytime cough counts of between1% and 99%. For instance, the objective daytime cough count can bereduced by between 50% and 90%, or cough count can be reduced by 75%. Inaddition, there may also be a significant reduction in daytime, andtotal 2 h cough count frequency, as well as cough severity score.

Administration and Pharmaceutical Composition

For example, the invention relates to a method for treating the symptomsof cough and urge to cough associated with a respiratory disease byadministering a compound of Formula (I). For example, the inventionrelates to a method of treatment of the symptoms of cough and/or urge tocough associated with a respiratory disease or disorder mediated by aP2X3 or P2X2/3 receptor antagonist by administering a compound ofFormula (I).

For example, the invention relates to methods for reducing daytime coughin idiopathic/treatment-resistant chronic cough. The invention alsorelates to a method of treating neuronal hypersensitivity underlyingchronic cough.

For example, the methods of the invention relate to treating, preventingor ameliorating the respiratory diseases and disorders described herein,or symptoms thereof, described herein in a patient in need thereof byadministering a compound selected from Compounds 1-39. For example, thecompound is selected from Compounds 6, 7, 13, 17, 21, 28, 35 and 38. Forexample, the compound is Compound 16.

The invention includes pharmaceutical compositions comprising at leastone compound of the present invention, or an individual isomer, racemicor non-racemic mixture of isomers or a pharmaceutically acceptable saltor solvate thereof, together with at least one pharmaceuticallyacceptable carrier, and optionally other therapeutic and/or prophylacticingredients.

In general, the compounds of the invention will be administered in atherapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. Suitable dosageranges are typically 1-1000 mg daily or twice daily, preferably 100-900mg daily or twice daily, and most preferably 500-700 mg daily or twicedaily, depending upon numerous factors such as the severity of thedisease to be treated, the age and relative health of the subject, thepotency of the compound used, the route and form of administration, theindication towards which the administration is directed, and thepreferences and experience of the medical practitioner involved.Suitable dosage ranges can also include dosages comprising 1-1000 mgmultiple times (e.g., 3-4 times) per day.

For instance, in some embodiments, the compounds of the invention can beadministered at a dosage of about 600 mg twice daily. In someembodiments, the compounds of the invention can be administered at adosage of about 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 550 mg, 650 mg,700 mg, 800 mg, 900 mg, or 1000 mg twice daily.

The duration of treatment can last for days, weeks, months or years. Insome embodiments, treatment (e.g., administration of a compound of thepresent invention or a pharmaceutically acceptable salt thereof) lastsfor two weeks. In some embodiments, treatment lasts one month. In someembodiments, treatment can proceed indefinitely. In some embodiments ofthe present invention, a subject can be treated with a compound ofFormula (I) at a dosage of 600 mg twice daily for two weeks.

One of ordinary skill in the art of treating such diseases will be able,without undue experimentation and in reliance upon personal knowledgeand the disclosure of this Application, to ascertain a therapeuticallyeffective amount of the compounds of the present invention for a givendisease.

Compounds of the invention may be administered as pharmaceuticalformulations including those suitable for oral (including buccal andsub-lingual), rectal, nasal, topical, pulmonary, vaginal, or parenteral(including intramuscular, intraarterial, intrathecal, subcutaneous andintravenous) administration or in a form suitable for administration byinhalation or insufflation. The preferred manner of administration isgenerally oral using a convenient daily dosage regimen which can beadjusted according to the degree of affliction.

A compound or compounds of the invention, together with one or moreconventional adjuvants, carriers, or diluents, may be placed into theform of pharmaceutical compositions and unit dosages. The pharmaceuticalcompositions and unit dosage forms may be comprised of conventionalingredients in conventional proportions, with or without additionalactive compounds or principles, and the unit dosage forms may containany suitable effective amount of the active ingredient commensurate withthe intended daily dosage range to be employed. The pharmaceuticalcompositions may be employed as solids, such as tablets or filledcapsules, semisolids, powders, sustained release formulations, orliquids such as solutions, suspensions, emulsions, elixirs, or filledcapsules for oral use; or in the form of suppositories for rectal orvaginal administration; or in the form of sterile injectable solutionsfor parenteral use. Formulations containing about one (1) milligram ofactive ingredient or, more broadly, about 0.01 to about one hundred(100) milligrams, per tablet, are accordingly suitable representativeunit dosage forms.

The compounds of the invention may be formulated in a wide variety oforal administration dosage forms. The pharmaceutical compositions anddosage forms may comprise a compound or compounds of the presentinvention or pharmaceutically acceptable salts thereof as the activecomponent. The pharmaceutically acceptable carriers may be either solidor liquid. Solid form preparations include powders, tablets, pills,capsules, cachets, suppositories, and dispersible granules. A solidcarrier may be one or more substances which may also act as diluents,flavouring agents, solubilizers, lubricants, suspending agents, binders,preservatives, tablet disintegrating agents, or an encapsulatingmaterial. In powders, the carrier generally is a finely divided solidwhich is a mixture with the finely divided active component. In tablets,the active component generally is mixed with the carrier having thenecessary binding capacity in suitable proportions and compacted in theshape and size desired. The powders and tablets preferably contain fromabout one (1) to about seventy (70) percent of the active compound.Suitable carriers include but are not limited to magnesium carbonate,magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch,gelatine, tragacanth, methylcellulose, sodium carboxymethylcellulose, alow melting wax, cocoa butter, and the like. The term “preparation” isintended to include the formulation of the active compound withencapsulating material as carrier, providing a capsule in which theactive component, with or without carriers, is surrounded by a carrier,which is in association with it. Similarly, cachets and lozenges areincluded. Tablets, powders, capsules, pills, cachets, and lozenges maybe as solid forms suitable for oral administration.

Other forms suitable for oral administration include liquid formpreparations including emulsions, syrups, elixirs, aqueous solutions,aqueous suspensions, or solid form preparations which are intended to beconverted shortly before use to liquid form preparations. Emulsions maybe prepared in solutions, for example, in aqueous propylene glycolsolutions or may contain emulsifying agents, for example, such aslecithin, sorbitan monooleate, or acacia. Aqueous solutions can beprepared by dissolving the active component in water and adding suitablecolorants, flavors, stabilizers, and thickening agents. Aqueoussuspensions can be prepared by dispersing the finely divided activecomponent in water with viscous material, such as natural or syntheticgums, resins, methylcellulose, sodium carboxymethylcellulose, and otherwell-known suspending agents. Solid form preparations include solutions,suspensions, and emulsions, and may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

The compounds of the invention may be formulated for topicaladministration to the epidermis as ointments, creams or lotions, or as atransdermal patch. Ointments and creams may, for example, be formulatedwith an aqueous or oily base with the addition of suitable thickeningand/or gelling agents. Lotions may be formulated with an aqueous or oilybase and will in general also containing one or more emulsifying agents,stabilizing agents, dispersing agents, suspending agents, thickeningagents, or coloring agents. Formulations suitable for topicaladministration in the mouth include lozenges comprising active agents ina flavored base, usually sucrose and acacia or tragacanth; pastillescomprising the active ingredient in an inert base such as gelatine andglycerine or sucrose and acacia; and mouthwashes comprising the activeingredient in a suitable liquid carrier.

The subject compounds may be formulated for nasal administration. Thesolutions or suspensions are applied directly to the nasal cavity byconventional means, for example, with a dropper, pipette or spray. Theformulations may be provided in a single or multidose form. In thelatter case of a dropper or pipette, this may be achieved by the patientadministering an appropriate, predetermined volume of the solution orsuspension. In the case of a spray, this may be achieved for example bymeans of a metering atomizing spray pump.

The compounds of the invention may be formulated for aerosoladministration, particularly to the respiratory tract and includingintranasal administration. The compound will generally have a smallparticle size for example of the order of five (5) microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization. The active ingredient is provided in a pressurizedpack with a suitable propellant such as a chlorofluorocarbon (CFC), forexample, dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane, or carbon dioxide or other suitable gas. Theaerosol may conveniently also contain a surfactant such as lecithin. Thedose of drug may be controlled by a metered valve. Alternatively theactive ingredients may be provided in a form of a dry powder, forexample a powder mix of the compound in a suitable powder base such aslactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidine (PVP). The powder carrier will form agel in the nasal cavity. The powder composition may be presented in unitdose form for example in capsules or cartridges of e.g., gelatine orblister packs from which the powder may be administered by means of aninhaler.

The pharmaceutical preparations may be in unit dosage forms. In suchform, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,oral tape or lozenge itself, or it can be the appropriate number of anyof these in packaged form.

Other suitable pharmaceutical carriers and their formulations aredescribed in Remington: The Science and Practice of Pharmacy 1995,edited by E. W. Martin, Mack Publishing Company, 19th edition, Easton,Pa. Representative pharmaceutical formulations containing a compound ofthe present invention are described below.

EXAMPLES

The following preparations and examples are given to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representativethereof.

Example 15-(5-Bromo-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

The synthetic procedure used in this Example is outlined in Scheme B.

Step 1. 2-Bromo-5-isopropyl-phenol

A solution of 3-isopropyl phenol (4.975 g, 36.5 mmol) in 37 mL of CCl₄was cooled to −20° C. Bromine (1.9 mL, 38.4 mmol) was dissolved in 5.0mL CCl₄ and added drop-wise at such a rate that the internal temperaturewas maintained below −10° C. The mixture was allowed to warm to roomtemperature. After 12 hours the mixture was taken up in 100 mL CH₂Cl₂,washed with H₂O and then with brine. The combined organics were driedover Na₂SO₄, filtered and concentrated in vacuo to give 8.663 g of a 1:1mixture of 2-bromo-5-isopropyl-phenol and 4-bromo-5-isopropyl phenol asa dark oil). These two isomers were inseparable and were used togetherin step 2 below.

Step 2. 1-Bromo-4-isopropyl-2-methoxy-benzene

To a mixture of 2-bromo-5-isopropyl-phenol and 4-bromo-5-isopropylphenol from step 1 (8.663 g, 40.3 mmol), K₂CO₃ (16.710 g, 120.9 mmol) in50 mL DMF, was added iodomethane (3.0 mL, 48.3 mmol) with mechanicalstirring. The mixture was warmed to 50° C. for 4 hours. After cooling toroom temperature 300 mL H₂O was added and the solution was extractedwith diethyl ether (Et₂O), washed with H₂O and washed with brine. Thecombined organics were dried over MgSO₄, filtered and concentrated invacuo to give 1-bromo-4-isopropyl-2-methoxy-benzene and1-bromo-2-isopropyl-4-methoxy-benzene (6.621 g, 72%) as a 1:1inseparable mixture in the form of a pale yellow oil. This mixture ofregioisomers was used directly in step 3 below.

Step 3. 5-Bromo-2-isopropyl-4-methoxy-benzaldehyde

To a solution of 1-bromo-4-isopropyl-2-methoxy-benzene and1-bromo-2-isopropyl-4-methoxy-benzene from step 2 (6.621 g, 28.9 mmol)in 100 mL 1,2 dichloroethane was added TiCl₄ (6.3 mL, 57.8 mmol) at 0°C. After 10 minutes, dichloromethoxymethane (Cl₂CHOMe) (2.6 mL, 28.9mmol) was added and the mixture was warmed to reflux. After 3 hours themixture was cooled poured over ice and acidified with 50 mL 2 M HCl. Theresulting slurry was extracted with CH₂Cl₂, and washed with brine. Thecombined organics were dried over MgSO₄, filtered and concentrated invacuo to give a dark-green oil. Purification via flash chromatography(96:4 hexane/ethyl acetate) afforded5-bromo-2-isopropyl-4-methoxy-benzaldehyde and5-bromo-4-isopropyl-2-methoxy-benzaldehyde (2.876 g, 39%, 6.621 g, 72%)as a 1:1 mixture of inseparable isomers in the form of an orange oil,which was used directly in step 4.

Step 4. 5-Bromo-2-isopropyl-4-methoxy-phenol

To a solution of 5-bromo-2-isopropyl-4-methoxy-benzaldehyde and5-bromo-4-isopropyl-2-methoxy-benzaldehyde from step 3 (2.87 g, 11.2mmol) in 25 mL CH₂Cl₂ was added mCPBA (2.31 g, 13.4 mmol). After 16hours the mixture was taken up in 150 ml CH₂Cl₂ and washed with satNaHCO₃, and then with brine. The combined organic layers were dried overNa₂SO₄, filtered and concentrated in vacuo to give an oil that was takenup in 50 mL MeOH and 30 mL 4M NaOH. After 2 hours the mixture wasevaporated, diluted with water and acidified to pH=1 with concentratedHCl. The mixture was extracted with ethyl acetate (3×100 mL) and washedwith 100 mL brine. The combined organics were dried over Na₂SO₄,filtered and evaporated to give a mixture of5-bromo-2-isopropyl-4-methoxy-phenol and2-bromo-5-isopropyl-4-methoxy-phenol as an orange residue. Theseregioisomers were separable by flash chromatography (gradient: hexane,7:3, 1:1 hexane/CH₂Cl₂) to afford 5-bromo-2-isopropyl-4-methoxy-phenol(0.929, 34%) as a yellow oil which was used in the following step, and2-bromo-5-isopropyl-4-methoxy-phenol (0.404 g, 15%) as a yellow solid.

Step 5. (5-Bromo-2-isopropyl-4-methoxy-phenoxy)-acetonitrile

To a mixture of 5-bromo-2-isopropyl-4-methoxy-phenol from step 4 (0.831g, 3.4 mmol) and K₂CO₃ (0.562 g, 4.1 mmol) in 17 mL dimethyl formamide(DMF) was added iodoacetonitrile (0.594 g, 3.6 mmol). The mixture waswarmed to 60° C. for 30 minutes and then allowed to cool to roomtemperature. After cooling to room temperature the mixture was taken upin 50 mL of H₂O and extracted with 1:1 toluene/ethyl acetate, washedwith H₂O and then with brine. The combined organic layers were driedover Na₂SO₄, filtered and concentrated in vacuo to give a crude solid.Purification via flash chromatography (1:1 hexane/CH₂Cl₂) afforded(5-bromo-2-isopropyl-4-methoxy-phenoxy)-acetonitrile (0.611 g, 63%) as awhile solid.

Step 6.2-(5-Bromo-2-isopropyl-4-methoxy-phenoxy)-3-methoxy-acrylonitrile

Sodium hydride (0.122 g, 5.0 mmol, 60% w/w) was washed with dry hexanesand evaporated under a stream of nitrogen. 10 mL THF was added and themixture was cooled to 0° C.(5-Bromo-2-isopropyl-4-methoxy-phenoxy)-acetonitrile (0.577 g, 2.03mmol) was added in portions. After 30 min ethyl formate (4.9 mL, 60.9mmol) was added and the solution was warmed to 80° C. After 4.5 hoursthe mixture was cooled and 5.0 mL iodomethane was added in one portion.After 16 hours the solution was quenched with H₂O, concentrated invacuo, extracted with ethyl acetate, washed with H₂O and then washedwith brine. The combined organic layers were dried over Na₂SO₄, filteredand concentrated in vacuo. Purification via flash chromatography (9:1hexane/ethyl acetate) afforded2-(5-bromo-2-isopropyl-4-methoxy-phenoxy)-3-methoxy-acrylonitrile (0.319g, 48%) as a white solid.

Step 7. 5-(5-Bromo-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a solution of2-(5-bromo-2-isopropyl-4-methoxy-phenoxy)-3-methoxy-acrylonitrile (0.282g, 0.9 mmol) and guanidine carbonate (0.078 g, 0.4 mmol) in 10.0 mLdimethyl sulfoxide (DMSO) was added sodium methoxide (1.0 mL, 1.0M inMeOH). The mixture was warmed to 120° C. The methanol was collected viaa short-path condenser. After 3 h the mixture was cooled andconcentrated in vacuo to give a crude oil. Purification via flashchromatography (95:5 CH₂Cl₂/MeOH) afforded 17 (0.246 g, 77%) as a pinksolid; Mass Spec M+H=352. The above procedure may be used with variousdifferent phenols in step 1 and/or substituted guanidines in step 7under essentially the same reaction conditions to produce additionalcompounds. Additional compounds made according to the procedure ofExample 1 are shown in Table 1.

Example 25-(2-Isopropyl-5-methanesulfonyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a mixture of 5-(2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine(0.32 g, 1.17 mmol), prepared according to Example 2, andmethanesulfonic anhydride (0.81 g, 4.67 mmol) was addedtrifluoromethanesulfonic acid (0.45 g, 3.00 mmol), and the mixture washeated at 80° C. for 16 hrs. The reaction mixture was poured into icewater, basified with saturated NaHCO₃ solution and extracted intodichloromethane, which was dried over Na₂SO₄, filtered and concentratedin vacuo. The residue was purified via flash chromatography on silicagel (3% CH₃OH in CH₂Cl₂ with 0.1% NH₄OH) gave5-(2-isopropyl-5-methanesulfonyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamineas a white solid (0.248 g, 90%; 0.107 g), MS (M+H): 353.

Example 35-(5-Iodo-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a solution of5-(2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.40 g, 1.44mmol) in glacial acetic acid (4 ml) at room temperature was added asolution of iodine monochloride (0.28 g, 1.76 mmol) in glacial aceticacid (4 ml). Water (6 ml) was also added, and the reaction was stirredfor 16 hours, after which another portion of iodine monochloride (0.4 g,2.47 mmol) in glacial acetic acid (4 ml) was added. The reaction mixturewas stirred for an additional hour at room temperature. The acidicmixture was basified with saturated NaHCO₃ solution and extracted intodichloromethane. The organic layer was dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified via flash chromatography(5% CH₃OH in CH₂CL₂ with 0.1% NH₄OH) to give5-(5-iodo-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine as beigecolored solid (0.536 g, 92%). M+H 400.

Example 45-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzonitrile

A mixture of5-(5-iodo-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.37 g,0.925 mmol) and CuCN (0.12 g, 1.39 mmol) in DMF (5 ml) was heated at120° C. for 3 hours. Water (100 ml) was added, and the precipitate wascollected. The residue was triturated with methanolic dichloromethane(10% CH₃OH in CH₂Cl₂ with 0.1% NH₄OH) to release the product from itscopper complex and filtered. The filtrate was concentrated and purifiedvia flash chromatography (3% CH₃OH in CH₂Cl₂ with 0.1% NH₄OH) to give5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzonitrile aswhite solid (0.12 g, 44%): M+H 300.

Example 5

1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanoneand1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-2-hydroxy-4-isopropyl-phenyl]-ethanone

5-(2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine in anhydrousdichloroethane (20 mL) was added to trifluoroacetic acid (0.06 mL, 0.77mmol), acetyl chloride (0.31 mL, 4.37 mmol), and aluminum trichloride(583 mg, 4.37 mmol). After stirring for 22 hours at room temperature,water (1.2 mL) was added to the reaction at 0° C. The mixture was driedusing anhydrous sodium sulfate and concentrated in vacuo. Aqueous sodiumhydroxide (0.2M, 10 mL) was added to the residue and the mixture washeated at 100° C. for 1 hour. After cooling, the reaction was extractedwith dichloromethane The dichloromethane layer was dried using anhydrousmagnesium sulfate, concentrated, and purified with silica gel columnchromatography eluting with 96/4/0.1 dichloromethane/methanol/ammoniumhydroxide to yield1-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanone(72 mg, 31%) as off-white solid, MS (M+H)=317. Also recovered was1-[5-(2,4-diamino-pyrimidin-5-yloxy)-2-hydroxy-4-isopropyl-phenyl]-ethanone(43 mg, 20%) as pale yellow solid, MS (M+H)=303.

Example 65-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzoic acid

To a suspension of5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzonitrile (50mg, 0.17 mmol, from Example 15) in ethanol (1 mL) was added sodiumhydroxide (174 mg, 4.34 mmol, dissolved in 1 mL water). After refluxingovernight, the reaction was cooled in an ice bath. Aqueous hydrochloricacid (3M) was added until the pH of the reaction was 7. The white solidprecipitate was collected, washed with small amounts of water anddichloromethane, and dried to yield5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzoic acid:(51 mg, 96%, MS (M+H)=319), which was converted to the hydrochloridesalt.

Example 75-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzamide

To 5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzonitrile(49 mg, 0.16 mmol, from Example 15) suspended in ethanol (1 mL) wasadded sodium hydroxide (64 mg, 1.60 mmol, dissolved in 1 mL water). Thereaction was heated at 110° C. for 5 hours, cooled, and washed withdichloromethane (25 mL). The dichloromethane layer was concentrated andpurified by preparatory TLC plates (92/8/0.5dichloromethane/methanol/ammonium hydroxide) to yield5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzamide aswhite solid (9 mg, 17%, MS (M+H)=318), which was converted to thehydrochloride salt.

Example 8[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ureaStep 1. 5-(5-Amino-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To 5-(2-isopropyl-4-methoxy-5-nitro-phenoxy)-pyrimidine-2,4-diamine (2.1g, 6.58 mmol) suspended in ethanol (150 mL) in a Parr bomb, was added10% palladium on charcoal (210 mg). After hydrogenation in the Parrhydrogenator overnight at 35 psi, the reaction was filtered throughcelite. The celite pad was washed with ethanol and ethyl acetate and thefiltrate was concentrated. Purification with silica gel columnchromatography (92/8/0.1 dichloromethane/methanol/ammonium hydroxide)gave 5-(5-amino-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine asa pale orange solid (468 mg, 25%, (M+H)⁺=290), which was converted tothe hydrochloride salt.

Step 2.[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-urea

To 5-(5-amino-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (314mg, 1.09 mmol) suspended in water (3 mL) was added acetic acid (0.25 mL,4.34 mmol). Once all solids had dissolved, sodium cyanate (71 mg, 1.09mmol, dissolved in 1.5 mL water) was added dropwise. After 30 minutes,the reaction was concentrated and purified with silica gel columnchromatography eluting with 92/8/0.1 dichloromethane/methanol/ammoniumhydroxide to yield[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-urea asan off-white solid (244 mg, 68%, M+H)⁺=333), which was converted to ahydrochloride salt:

Example 9N-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-acetamide

To 5-(5-amino-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (100mg, 0.35 mmol, from Example 17) dissolved in anhydrous dichloromethane(10 mL) was added anhydrous pyridine (0.03 mL, 0.38 mmol). To thisreaction mixture at 0° C. was added acetyl chloride (0.03 mL, 0.38mmol). After stirring at room temperature for 1 hour, the reaction wasconcentrated and purified with preparatory TLC (93/7/0.5dichloromethane/methanol/ammonium hydroxide) to yield an off-white solid(74 mg mixture of bis- and tris-acetylated products). To this solid wasadded aqueous sodium hydroxide (0.2 M, 2 mL), and the mixture wasrefluxed for 1 hour, cooled, and washed with dichloromethane (10 mL).The dichloromethane layer was dried using anhydrous magnesium sulfateand concentrated in vacuo to yieldN-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-acetamideas a white solid (53 mg, 46%, M+H)⁺=332) which was converted to ahydrochloride salt:

Example 105-(2-Isopropyl-4-methoxy-5-nitro-phenoxy)-pyrimidine-2,4-diamine

The synthetic procedure used in this Example is outlined in Scheme D

Step 1. 2-(1-Hydroxy-1-methyl-ethyl)-4-methoxy-phenol

To a solution of methylmagnesium bromide (221 ml, 665 mmol) in 800 mlTHF at 0° C. was added 1-(2-hydroxy-5-methoxy-phenyl)-ethanone (20.21 g,302 mmol) in portions over 30 min. The mixture was allowed to warm toroom temperature. After 16 h the mixture was quenched by the slowaddition of 10% NH₄Cl, carefully acidified to pH=1 (slow addition) withconcentrated HCl and extracted with Et₂O. The combined organics werewashed with H₂O, washed with brine, died over MgSO₄, filtered andconcentrated in vacuo to give2-(1-hydroxy-1-methyl-ethyl)-4-methoxy-phenol (50.57 g, 100%) as a tansolid.

Step 2. 2-Isopropyl-4-methoxy-phenol

To a solution of 2-(1-hydroxy-1-methyl-ethyl)-4-methoxy-phenol (50.57 g,278 mmol) in 550 ml AcOH was added 10% Pd/C (as a slurry in 20 ml H₂O).Ammonium formate (87.52 g, 1388 mmol) was added in portions. The mixturewas warmed to 100° C. for 1 hour, cooled and filtered through a pad ofcelite. The celite pad was washed with ethyl acetate. The mother liquorwas mixed with H₂O and extracted with ethyl acetate. The combinedorganics were washed with H₂O, washed with brine, dried over Na₂SO₄,filtered and concentrated in vacuo to give 2-isopropyl-4-methoxy-phenol(44.74 g, 97%) as a pale yellow oil.

Step 3. Toluene-4-sulfonic acid 2-isopropyl-4-methoxy-phenyl ester

To a solution of 2-isopropyl-4-methoxy-phenol (56.91 g, 342 mmol)triethylamine (57.3.0 ml, 411 mmol) in 750 ml CH₂Cl₂ was cooled to 0° C.p-Toluenesulfonyl chloride (68.54 g, 360 mmol) in 250 ml CH₂Cl₂ wasadded drop-wise at a rate that maintained the internal temperature <10°C. The mixture was allowed to warm to rt. After 16 h, H₂O was added andthe mixture was extracted with CH₂Cl₂. The combined organics were washedwith brine, dried with Na₂SO₄, filtered and concentrated in vacuo toafford a crude solid. Recrystallization from hexanes affordedtoluene-4-sulfonic acid 2-isopropyl-4-methoxy-phenyl ester (81.67 g,74%) as white needles.

Step 4. Toluene-4-sulfonic acid 2-isopropyl-4-methoxy-5-nitro-phenylester

To a solution of toluene-4-sulfonic acid 2-isopropyl-4-methoxy-phenylester (19.00 g, 59 mmol) in 118 mL AcOH was added 236 ml fuming HNO₃over 20 min. After 16 h the solution was pouring into a rapidly stirringslurry of 21 of ice/H₂O. After 15 min the precipitate was filtered,washed with H₂O and dried under vacuum (50° C.) to givetoluene-4-sulfonic acid 2-isopropyl-4-methoxy-5-nitro-phenyl ester(21.27 g, 98%) and toluene-4-sulfonic acid2-isopropyl-4-methoxy-3-nitro-phenyl ester and as a pale yellow solid(7:1 inseparable mixture).

Step 5. 2-Isopropyl-4-methoxy-5-nitro-phenol

A solution of toluene-4-sulfonic acid2-isopropyl-4-methoxy-5-nitro-phenyl ester and2-isopropyl-4-methoxy-3-nitro-phenyl ester (21.20 g, 58 mmol) and 175 mL2M KOH in 350 mL EtOH was warmed to 100° C. After 45 minutes the mixturewas cooled, evaporated and taken up in 1 l of water. The solution wasacidified to pH=1 with 12 M HCl and extracted with ethyl acetate. Thecombined organics were washed with H₂O, brine, dried over Na₂SO₄,filtered and concentrated in vacuo. The crude oil was purified via flashchromatography (gradient: 95:5 to 4:1 hexane/ethyl acetate) to afford3-amino-2-isopropyl-5-nitro-phenol (10.03 g, 81%) as a yellow solid and3-amino-2-isopropyl-3-nitro-phenol (1.32 g, 11%) as a yellow oil.

Step 6. (2-Isopropyl-4-methoxy-5-nitro-phenoxy)-acetonitrile

A mixture of 3-amino-2-isopropyl-5-nitrophenol (9.94 g, 47 mmol), K₂CO₃(13.00 g, 94 mmol) and toluenesulfonic acid cyanomethyl ester (10.93 g,52 mmol) in 500 mL DMF was warmed to 50° C. After 16 h the mixture wascooled, poured into 500 mL H₂O and extracted with toluene/ethyl acetate(1:1). The combined organics were washed with H₂O, washed with brine,filtered and concentrated in vacuo. The crude solid was recrystallizedfrom EtOH to afford (2-isopropyl-4-methoxy-5-nitro-phenoxy)-acetonitrile(8.95 g, 76%) as a yellow crystalline solid.

Step 7. 5-(2-Isopropyl-4-methoxy-5-nitro-phenoxy)-pyrimidine-2,4-diamine

A mixture of (2-isopropyl-4-methoxy-5-nitro-phenoxy)-acetonitrile (8.785g, 35.5 mmol) and Bredereck's reagent (14.6 mL, 70.9 mmol) was warmed to100° C. After 45 min the mixture was evaporated under reduced pressure(50° C., 50 mtorr) to give an orange solid. The solid was added to asolution of aniline hydrochloride (9.19 g, 70.9 mmol) in 150 mL of EtOH.The mixture was warmed to reflux. After 16 hr additional anilinehydrochloride (4.596 g, 35.5 mmol) was added mixture was continued atreflux for 4 h. The solution was concentrated in vacuo and poured intoH₂O. The mixture was extracted with ethyl acetate, washed with H₂O,washed with brine, dried over Na₂SO₄, and concentrated in vacuo toafford a yellow-green solid. This crude product was added to a mixtureof 200 mL NMP and guanidine carbonate (17.70 g, 98 mmol) and warmed to130° C. After 5 hours the mixture was cooled then poured onto 21 of anice/H₂O mixture. The resulting precipitate was filtered, washed with H₂Oand dried under vacuum (50° C.). The crude solid was recrystallized fromEtOH to afford5-(2-isopropyl-4-methoxy-5-nitro-phenoxy)-pyrimidine-2,4-diamine (8.14g, 63%, 3 steps) as a yellow crystalline solid (solvated 1:1 with EtOH).(M+H)⁺=320.

Example 111-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-3-ethyl-ureaStep 1. 5-(5-Amino-2-Isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a solution of5-(2-isopropyl-4-methoxy-5-nitro-phenoxy)-pyrimidine-2,4-diamine (2.953g, 9.2 mmol) in 250 mL EtOH and 25 AcOH was added 10% Pd/C. The mixturewas placed under 50 psi of H₂ via a Parr hydrogenator. After 2.5 h themixture was filtered through a pad of celite. The pad was washed withethyl acetate and the solution was partially concentrated in vacuo. Theresidue was taken up in 500 mL H₂O and cooled to 0° C. The solution wasadjusted to pH=12 with 50% NaOH extracted with ethyl acetate. Thecombined organics were washed with H₂O, washed with brine, dried overNa₂SO₄, filtered and concentrated in vacuo to afford5-(5-amino-2-Isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (2.156g, 82%) as a dark-orange solid.

Step 2.1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-3-ethyl-urea

A solution of5-(5-amino-2-Isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.117g, 0.4 mmol) and ethyl isocyanate (0.034 g, 0.5 mmol) in 4 mL of toluenewas heated to 100° C. in a sealed tube. After 5 h the solution wascooled and concentrated in vacuo gave a brown solid. Purification viaflash chromatography (CH₂Cl₂/MeOH 97:3) afforded1-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-3-ethyl-urea(0.120 g, 83%) as a white solid; (M+H)=361.

Example 121-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-3-phenyl-urea

5-(5-amino-2-Isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.309g, 1.1 mmol) was converted, as described in the above procedure, to1-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-3-phenyl-urea(0.122 g, 28%) as white solid; [MH]⁺=408.

Similarly prepared from5-(5-amino-2-Isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.313g, 1.1 mmol) and 2,5-hexanedione (0.14 ml, 1.2 mmol) was5-[5-(2,5-Dimethyl-pyrrol-1-yl)-2-isopropyl-4-methoxy-phenoxy]-pyrimidine-2,4-diamine,(0.259 g, 64%). (M+H)=368.

Example 134-Chloro-N-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-butyramide

To a solution of5-(5-amino-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.400g, 1.4 mmol) in 15 ml CHCl₃ and Na₂HPO₄ (0.392 g, 2.8 mmol) was added4-chlorobutyryl chloride (0.194 g, 1.4 mmol) drop-wise. After 4.5 h, H₂Oand CH₂Cl₂ were added and the mixture was allowed to stir 15 min. Themixture was neutralized with 2N Na₂CO₃ and extracted with CH₂Cl₂. Thecombined organics were washed with brine, dried over Na₂SO₄, filteredand concentrated in vacuo to afford4-chloro-N-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-butyramide(0.495 g, 91%) as brown foam; [MH]⁺=394.

Example 145-(2-Isopropyl-5-isothiocyanato-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a solution of5-(5-amino-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.100g, 0.4 mmol) in 1 ml H₂O and TFA (0.040 g, 0.4 mmol) was addedthiophosgene (0.040 g, 0.4 mmol). After 1 h the mixture was neutralizedwith 2M NaOH and extracted with CH₂Cl₂. The combined organics werewashed with brine, dried over Na₂SO₄, filtered and concentrated in vacuoto afford5-(2-isopropyl-5-isothiocyanato-4-methoxy-phenoxy)-pyrimidine-2,4-diamine(0.042 g, 36%) as brown foam [MH]⁺=334.

Example 152-[5-(2,4-Diaminopyrimidin-5-yloxy)-4-isopropyl-2methoxy-phenyl]-propan-2-ol

To a solution of methylmagnesium bromide (83.4 mmol, 27.8 ml, 3.0 M inEt₂O) in 83 mL THF at 0° C. was added1-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanone(2.523 g, 8.3 mmol, from Example 16) in portions. After 16 h the mixturewas cooled to 0° C. and was quenched by the addition 10% NH₄Cl. H₂O wasadded and the mixture was extracted with ethyl acetate. The combinedorganics were washed with H₂O, washed with brine, dried over NaHCO₃,filtered and concentrated in vacuo. The crude solid was taken up in 31ml DMF. K₂CO₃ (0.65 g, 4.7 mmol) and iodomethane (0.098 ml, 1.6 mmol)were added and the mixture was warmed to 50° C. Additional portions ofiodomethane (0.019 mL, 0.6 mmol) was added at 1, 2 and 3 hr. After 16 hthe mixture was cooled and 10% NH₄Cl and extracted with ethyl acetate.The combined organics were washed with H₂O, washed with brine, driedwith Na₂SO₄, filtered and concentrated in vacuo to give2-[5-(2,4-diaminopyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-propan-2-ol(0.711 g, yield) as a white solid. [MH]⁺=333.

Example 16 5-(2,5-Diiosopropyl-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a solution of2-[5-(2,4-diaminopyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-propan-2-ol:(0.350 g, 1.1 mmol) in 10 ml CH₂Cl₂ was added trifluoroacetic acid (4.0ml, 52.6 mmol) and triethylsilane (1.7 ml, 10.5 mmol). After 30 minsaturated NaHCO₃ was added and the mixture was extracted with ethylacetate. The combined organics were washed with brine, dried overNa₂SO₄, filtered and concentrated in vacuo to give a crude oil.Purification via flash chromatography (96:4 CH₂Cl₂/MeOH) gave5-(2,5-diiosopropyl-methoxy-phenoxy)-pyrimidine-2,4-diamine (0.225 g,68%) as a white solid. [MH]⁺=317.

Example 171-[5-(2,4-Diamino-pyrimidine-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanol

To a solution of1-[5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanone(2.500 g, 8.3 mmol) in 100 ml MeOH was slowly added NaBH₄(1.566 g, 41.4mmol) at 0° C. The solution was allowed to warm to rt. After 20 h, thesaturated NH₄Cl was added, the mixture was concentrated in vacuo andextracted with ethyl acetate. The combined organics were washed withbrine, dried over Na₂SO₄, filtered and concentrated in vacuo.Purification via silica gel column chromatography (9:1 CH₂Cl₂/MeOH)afforded to1-[5-(2,4-diamino-pyrimidine-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanol(1.613 g, 60%) as white foam; [MH]⁺=301.

Example 185-(2-Isopropyl-4-methoxy-5-vinyl-phenoxy)-pyrimidine-2,4-diamine and5-[2-Isopropyl-4-methoxy-5-(1-methoxy-ethyl)-phenoxy]-pyrimidine-2,4-diamine

To a solution of1-[5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-phenyl]-ethanol(1.613 g, 5.3 mmol) in 30 ml CH₂Cl₂ at −78° C. was added DAST (0.935 g,5.8 mmol). After stirring 1.5 h, saturated NaHCO₃ was added and themixture was extracted by CH₂Cl₂. The combined organics were washed withbrine and dried with Na₂SO₄, filtered and concentrated in vacuo.Purification via silica gel chromatography (95:5 CH₂Cl₂/MeOH) gave5-(2-Isopropyl-4-methoxy-5-vinyl-phenoxy)-pyrimidine-2,4-diamine (0.044g, 3%) as a foam ([MH]⁺=301) and5-[2-Isopropyl-4-methoxy-5-(1-methoxy-ethyl)-phenoxy]-pyrimidine-2,4-diamine(0.075g, 4%) as foam. [MH]⁺=303.

Example 195-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-N-methyl-benzenemethylsulfonamideStep 1.5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzenesulfonylchloride

A mixture of pyrimidine (0.400 g, 1.5 mmol) in 2 ml chlorosulfonic acidwas allowed to stir 20 min. The mixture was poured over ice. Theprecipitate was filtered, washed by cold H₂O and dried under vacuum toafford5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzenesulfonylchloride (0.515 g, 95%) as a white solid; [MH]⁺=373.

Step 2.5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-N-methyl-benzenemethylsulfonamide

To 10 ml methyl amine −78° C. in a screw-capped tube was added5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzenesulfonylchloride (0.300 g, 0.8 mmol). The mixture was allowed to warm to roomtemperature. After 20 hours the mixture was evaporated, washed with H₂O,and dried under vacuum to afford5-(2,4-diamino-pyrimidin-5-yloxy)-4-idopropyl-2-methoxy-N-methyl-benzenemethylsulfonamide(0.170 g, 57%) as a white solid; mp (HCl salt)=252.3-252.9° C.;[MH]⁺=367.

Similarly prepared, replacing methylamine with ethylamine, was5-(2,4-Diamino-pyrimidin-5-yloxy)-N-ethyl-4-isopropyl-2-methoxy-benzenesulsonamide(0.186 g, 61%) as a white solid; mp (HCl salt)=260-265° C.; [MH]⁺=382.

Example 205-(2,4-Diamino-pyrimidin-5-yloxy)4-isopropyl-2-methoxy-N,N-dimethyl-benzamide

To a suspension of5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzoic acid(180 mg, 0.57 mmol, from Example 17) in anhydrous dichloromethane (5.6mL) was added trifluoroacetic acid (0.08 mL, 1.14 mmol) and then thionylchloride (0.36 mL, 5.65 mmol). After 1 hour the reaction wasconcentrated. To the residue was added anhydrous dichloromethane (4.5mL) and dimethylamine (2.84 mL of a 2M solution in tetrahydrofuran, 5.65mmol). After 2 hours stirring at room temperature, the reaction wasfiltered and concentrated. Purification via silica gel columnchromatography eluting with 95/5/0.1 to 93/7/0.1dichloromethane/methanol/ammonium hydroxide yielded5-(2,4-diamino-pyrimidin-5-yloxy)4-isopropyl-2-methoxy-N,N-dimethyl-benzamide(40 mg, 20%) as pale yellow solid, MS (M+H)=346.

Similarly prepared using methylamine instead of dimethylamine,5-(2,4-diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-N-methyl-benzamide(23 mg, 15%) was prepared as pale yellow solid, MS (M+H)=332.

Example 21 4-(2,4-Diamino-pyrimidin-5-yloxy)-2-iodo-5-isopropyl-phenol

To a cold suspension of 1(0.21 g, 0.52 mmol) in dichloromethane (15 ml)at 0° C. was added BBr₃(0.26 g, 1.05 mmol). The reaction mixture wasstirred at room temperature for 16 hrs., quenched with water andbasified with sat. NaHCO₃. The insoluble solid was collected byfiltration. The filtrate was washed with water, dried over Na₂SO₄,filtered and concentrated in vacuo. The combined residue was purifiedvia flash chromatographed on silica gel (3 to 5% methanol indichloromethane with 0.1% NH₄OH) gave desired product (0.174 g, 86%),(M+H)=387.

Example 225-(5-Iodo-2-isopropyl-4-prop-2-ynyloxy-phenoxy)-pyrimidine-2,4-diamine

To 4-(2,4-Diamino-pyrimidin-5-yloxy)-2-iodo-5-isopropyl-phenol (200 mg,0.43 mmol) dissolved in anhydrous N,N-dimethylformamide (2 mL) was addedanhydrous potassium carbonate (414 mg, 3.00 mmol) and propargyl chloride(0.03 mL, 0.43 mmol). After stirring at room temperature overnight, thereaction was extracted with dichloromethane, water and brine. Thedichloromethane layer was dried using anhydrous magnesium sulfate,concentrated, and purified via silica gel column chromatography(95/5/0.1 dichloromethane/methanol/ammonium hydroxide) to yield5-(5-iodo-2-isopropyl-4-prop-2-ynyloxy-phenoxy)-pyrimidine-2,4-diamineas white solid (131 mg, 71%), MS (M+H)=425.

Example 235-(5-Ethanesulfonyl-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine

To a solution of sodium sulfite (541 mg, 4.29 mmol) in water (20 mL) wasadded5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzenesulfonylchloride (400 mg, 1.07 mmol) and the reaction was heated at 80° C. for 1hour. Sodium bicarbonate (361 mg, 4.29 mmol-dissolved in 5 mL water),dioxane (20 mL), and ethyl iodide (0.10 mL, 1.29 mmol) were added andthe reaction was heated at 80° C. for 2 hours. The reaction wasconcentrated, extracted with dichloromethane (150 mL) and water (20 mL).The dichloromethane layer was dried using anhydrous sodium sulfate,concentrated, and purified via silica gel column chromatography(95/5/0.1 dichloromethane/methanol/ammonium hydroxide) to yield5-(5-ethanesulfonyl-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine(77 mg, 20%) as white solid, MS (M+H)=367.

Example 245-(2-Isopropyl-4-methoxy-5-trifluoromethyl-phenoxy)-pyrimidine-2,4-diamine

The synthetic procedure used in this Example is outlined in Scheme E.

Step 1. 1-Iodo-4-isopropyl-2-methoxy-5-(toluene-4-sulfonyl)-benzene

To a solution of 2-Isopropyl-4-methoxy-1-(toluene-4-sulfonyl)-benzene(10 g, 31.25 mmol) in HOAc (10 ml) was added a solution of ICl (9.6 g,59.26 mmol) in HOAc (10 ml) and H₂O (5 ml). The reaction mixture wasstirred at room temperature for 16 hrs and basified by saturated NaHCO₃solution. The aqueous solution was extracted into EtOAc which was washedwith water, brine, dried over Na₂SO₄, filtered and concentrated in vacuoto give 1-Iodo-4-isopropyl-2-methoxy-5-(toluene-4-sulfonyl)-benzene(12.35 g, 89%).

Step 2.1-Isopropyl-5-methoxy-2-(toluene-4-sulfonyl)-4-trifluoromethyl-benzene

To a hot mixture of1-Iodo-4-isopropyl-2-methoxy-5-(toluene-4-sulfonyl)-benzene (0.5 g, 1.12mmol), CuI, KF in anhydrous DMF (10 ml) at 120° C. oil bath temperature,was added trifluoromethyl iodide (0.64 g, 4.48 mmol) in portions over 30min. The reaction mixture was heated for 4 hrs and poured into H₂O (100ml). The insoluble solid, which was collected by filtration, wastriturated with methylene chloride, filtered and concentrated to give1-Isopropyl-5-methoxy-2-(toluene-4-sulfonyl)-4-trifluoromethyl-benzene(0.45 g, 100%) as a solid.

Step 3. 2-Isopropyl-4-methoxy-5-trifluoromethyl-phenol

A solution of1-Isopropyl-5-methoxy-2-(toluene-4-sulfonyl)-4-trifluoromethyl-benzene(0.40 g, 1.03 mmol) and NaOH (0.5 g, 12.5 mmol) in MeOH(5 ml) and H₂O (5ml) was heated at 90° C. for 2 hrs. The cooled reaction mixture wasacidified with 3N HCl and extracted into methylene chloride. Thecombined extracts was dried with Na₂SO₄, filtered and concentrated togive desired 2-Isopropyl-4-methoxy-5-trifluoromethyl-phenol (0.194 g,81%) as an oil.

Step 4.5-(2-Isopropyl-4-methoxy-5-trifluoromethyl-phenoxy)-pyrimidine-2,4-diamine

Following the procedure of Example 2 steps 5-7,2-Isopropyl-4-methoxy-5-trifluoromethyl-phenol was converted to5-(2-Isopropyl-4-methoxy-5-trifluoromethyl-phenoxy)-pyrimidine-2,4-diamine.(M+H)=343

Example 255-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-thiobenzamide

A mixture of5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-benzamide (0.25g, 0.79 mmol, prepared according to the procedure of Example 52) andLawesson's reagent (0.96 g, 2.37 mmol) in anhydrous THF (20 ml) wasstirred at room temperature for 16 hrs and concentrated in vacuo. Flashchromatography on silica (5% CH₃OH in methylene chloride with 1% NH₄OH)gave5-(2,4-Diamino-pyrimidin-5-yloxy)-4-isopropyl-2-methoxy-thiobenzamide(0.201 g, 76%) as a yellow solid.

Example 265-(4-Ethoxy-5-iodo-2-isopropyl-phenoxy)-pyrimidine-2,4-diamine

To a solution of4-(2,4-Diamino-pyrimidin-5-yloxy)-2-iodo-5-isopropyl-phenol (0.2 g, 0.52mmol) in anhydrous DMF (2 ml) was added EtBr (57 mg, 0.52 mmol) inportions. The reaction mixture was partitioned between EtOAc and H₂O.The organic extract was dried over Na₂SO₄, filtered and concentrated.Flash chromatography on silica (3% MeOH in methylene chloride with 1%NH₄OH) gave5-(4-Ethoxy-5-iodo-2-isopropyl-phenoxy)-pyrimidine-2,4-diamine (0.17 g,28%) as a yellow solid. (M+H)=415.

Example 27

Formulations

Pharmaceutical preparations for delivery by various routes areformulated as shown in the following Tables. “Active ingredient” or“Active compound” as used in the Tables means one or more of theCompounds of Formula (I).

Composition for Oral Administration

Ingredient % wt./wt. Active ingredient 20.0% Lactose 79.5% Magnesiumstearate 0.5%

The ingredients are mixed and dispensed into capsules containing about100 mg each; one capsule would approximate a total daily dosage.

Composition for Oral Administration

Ingredient % wt./wt. Active ingredient 20.0% Magnesium stearate 0.5%Crosscarmellose sodium 2.0% Lactose 76.5% PVP (polyvinylpyrrolidine)1.0%

The ingredients are combined and granulated using a solvent such asmethanol. The formulation is then dried and formed into tablets(containing about 20 mg of active compound) with an appropriate tabletmachine.

Composition for Oral Administration

Ingredient Amount Active compound 1.0 g Fumaric acid 0.5 g Sodiumchloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 g Granulatedsugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum K (Vanderbilt Co.)1.0 g Flavoring 0.035 ml Colorings 0.5 mg Distilled water q.s. to 100 ml

The ingredients are mixed to form a suspension for oral administration.

Topical Formulation

Ingredients Grams Active compound 0.2-2 Span 60 2 Tween 60 2 Mineral oil5 Petrolatum 10 Methyl paraben 0.15 Propyl paraben 0.05 BHA (butylatedhydroxy anisole) 0.01 Water q.s. 100

All of the ingredients, except water, are combined and heated to about60° C. with stirring. A sufficient quantity of water at about 60° C. isthen added with vigorous stirring to emulsify the ingredients, and waterthen added q.s. about 100 g.

Nasal Spray Formulations

Several aqueous suspensions containing from about 0.025-0.5 percentactive compound are prepared as nasal spray formulations. Theformulations optionally contain inactive ingredients such as, forexample, microcrystalline cellulose, sodium carboxymethylcellulose,dextrose, and the like. Hydrochloric acid may be added to adjust pH. Thenasal spray formulations may be delivered via a nasal spray metered pumptypically delivering about 50-100 microliters of formulation peractuation. A typical dosing schedule is 2-4 sprays every 4-12 hours.

Compound 16 Tablets

Compound 16 is supplied formulated in a yellow, film-coated, oval-shapedtablet containing 10, 20, 30, 50, 100 or 300 mg of Compound 16. Tabletsare formulated with USP/NF compendial grade lactose monohydrate,hydroxypropyl methyl cellulose (HPMC or Hypromellose), croscarmellosesodium, microcrystalline cellulose (Avicel PH102), and magnesiumstearate as described in Table 2. Tablets are film-coated with OpadryYellow (Colorcon, Inc.) and packaged in HDPE bottles with childresistant caps and induction seals.

TABLE 2 Compound 16: Quantitative Tablet Composition (300 mg andPlacebo) Amount Amount for for 300 mg Placebo Tablet Tablet ComponentGrade Function (mg) (mg) Intragranular Compound 16 In house Active 300.00 (milled) Lactose USP/NF Diluent 187.8 487.8 monohydrate CroscarmelloseUSP/NF Disintegrant 18.0 18.0 sodium Hydroxypropyl USP/NF Binder 18.018.0 methyl cellulose Extragranular Croscarmellose USP/NF Disintegrant12.0 12.0 sodium Microcrystalline USP/NF Diluent 60.0 60.0 CelluloseMagnesium Stearate USP/NF Lubricant 4.2 4.2 Core Tablet 600 600 FilmCoating Opadry Yellow * Film-coat 18.0 18.0 03K12429 Sterile Water forUSP/NF Granulating As needed As needed Irrigation Solution Total Weightof Film 618 618 Coated Tablet * Opadry Yellow is composed of thefollowing USP/NF excipients: hypromellose, titanium dioxide, talc,triacetin and yellow iron oxide.

Example 28 P2X3/P2X2/3 FLIPR (Fluorometric Imaging Plate Reader) Assay

CHO-K1 cells were transfected with cloned rat P2X3 or human P2X2/3receptor subunits and passaged in flasks. 18-24 hours before the FLIPRexperiment, cells were released from their flasks, centrifuged, andresuspended in nutrient medium at 2.5×10⁵ cells/ml. The cells werealiquoted into black-walled 96-well plates at a density of 50,000cells/well and incubated overnight in 5% CO₂ at 37° C. On the day of theexperiment, cells were washed in FLIPR buffer (calcium- andmagnesium-free Hank's balanced salt solution, 10 mM HEPES, 2 mM CaCl₂,2.5 mM probenecid; FB). Each well received 100 μl FB and 100 μl of thefluorescent dye Fluo-3 AM [2 μM final conc.]. After a 1 hour dye loadingincubation at 37° C., the cells were washed 4 times with FB, and a final75 μl/well FB was left in each well.

Test compounds (dissolved in DMSO at 10 mM and serially diluted with FB)or vehicle were added to each well (25 μl of a 4X solution) and allowedto equilibrate for 20 minutes at room temperature. The plates were thenplaced in the FLIPR and a baseline fluorescence measurement (excitationat 488 nm and emission at 510-570 nm) was obtained for 10 seconds beforea 100 μl/well agonist or vehicle addition. The agonist was a 2× solutionof α,β-meATP producing a final concentration of 1 μM (P2X3) or 5 μM(P2X2/3). Fluorescence was measured for an additional 2 minutes at 1second intervals after agonist addition. A final addition of ionomycin(5 μM, final concentration) was made to each well of the FLIPR testplate to establish cell viability and maximum fluorescence of dye-boundcytosolic calcium. Peak fluorescence in response to the addition ofα,β-meATP (in the absence and presence of test compounds) was measuredand inhibition curves generated using nonlinear regression. PPADS, astandard P2X antagonist, was used as a positive control.

Using the above procedure, compounds of the invention exhibited activityfor the P2X3 receptor. The compound4-(2,4-Diamino-pyrimidin-5-yloxy)-2-iodo-5-isopropyl-phenol, forexample, exhibited a pIC₅₀ of approximately 8.3 using the above assay.

Example 29 In vivo Assay for Cough Sensitization

Hartley guinea pigs are studied in a standard tussive protocol. Briefly,guinea pigs (N=4-8/group) are treated with inhalation of nebulizedcitric acid following prior sensitization with inhaled histamine oralpha, beta-methylene ATP, and monitored for the development of coughingnots, as observed by an experienced investigator. Animals receivevehicle (p.o. or by nebulized inhalation) or a compound of the invention(from 1 to 100 mg/kg p.o., or at increasing nebulized concentrations),30-60 minutes prior to to the inhalational challenge of histamine oralpha, beta-methylene ATP, followed by citric acid solution.

Cough responsiveness is then monitored by counting frequency followingchallenge provocation, such that the magnitude by which compound of theinvention inhibits the frequency of tussive response can be calculated.

Example 30 Inhibition of ATP-gated P2X3 channels by Compound 16: anEffective Anti-Tussive Mechanism in Chronic Cough

Evidence suggests that P2X3 receptors are expressed by airway vagalafferents and contribute to the hyperexcitability of sensory neurons.Thus, the inventors explored the role of P2X3 receptors in thesensitization of vagal pathways mediating the cough reflex leading tochronic cough (CC). A cough A studs' was performed to investigate theefficacy of oral administration of Compound 16 in reducing daytime coughin idiopathic/treatment-resistant chronic cough.

In this study, a double-blind randomized placebo-controlled crossovertrial, 24 subjects (19 women, mean age 55 yrs) were randomized into adouble blind, placebo-controlled, 2-period, crossover study, of Compound16, 600 mg bd. Cough was assessed using an ambulatory sound monitoringsystem at baseline and after 2 weeks of treatment: primary endpoint,daytime objective cough frequency (coughs/hr) (VitaloJAIKJ™): secondaryendpoints, cough severity and urge to cough (UTC) visual analogue scales(VAS), global ratings of change and cough quality of life questionnaire(CQLQ).

Compound 16 markedly reduced cough (mean difference vs. placebo):daytime coughs/hr −75% (95% Cl −50 to −88), p<0.001 (see the FIGURE):cough severity VAS −26mm (−10 to −42), p=0.003: UTC VAS −21mm (−2 to−41), p=0.035; and CQLQ −9(−2 to −17). p=0.018. 13/24 of Compound 16patients rated an improved cough compared to 2/22 treated with placebo.Therefore, Compound 16 demonstrated substantial anti-tussive effectsrelative to placebo. In fact, treatment with Compound 16surprisinglyproduced a 75% reduction in objective daytime cough counts, in patientswith refractory chronic cough.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

What is claimed is:
 1. A method for treating a cough in a patient, wherein the cough is selected from the group consisting of chronic pathological cough, sub-acute pathological cough, acute pathological cough, neuronal hypersensitivity underlying chronic cough, neuronal hypersensitivity underlying sub-acute cough and neuronal hypersensitivity underlying acute cough, the method comprising administering to the patient in need thereof an effective amount of a compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is hydrogen or C₁-C₆ alkyl, wherein C₁-C₆ alkyl is optionally substituted with at least one substituent selected from the group consisting of cycloalkyl, halo, nitro, cyano, hydroxy, alkoxy, amino, acylamino, monoalkylamino, dialkylamino, haloalkoxy, heteroalkyl, —COR, —(CR′R″)_(n)—COOR and —(CR′R″)_(n)—CONR^(a)R^(b), wherein: in COR, R is hydrogen, alkyl, phenyl or phenylalkyl; in (CR′R″)_(n)—COOR, R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl; each n is independently 0, 1, 2, 3, 4 or 5; each R′ is independently hydrogen or alkyl; each R″ is independently hydrogen or alkyl; R^(a) is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl; R^(b) is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl; and R² R is alkyl, alkenyl, alkynyl, amino, aminosulfonyl, halo, amido, haloalkyl, alkoxy, hydroxy, haloalkoxy, nitro, hydroxyalkyl, alkoxyalkyl, hydroxyalkoxy, alkynylalkoxy, alkylsulfonyl, arylsulfonyl, carboxyalkyl, cyano or alkylcarbonyl.
 2. The method of claim 1, wherein the cough is chronic pathological cough.
 3. The method of claim 1, wherein the cough is neuronal hypersensitivity underlying acute cough.
 4. The method of claim 1, wherein the cough is selected from the group consisting of neuronal hypersensitivity underlying chronic cough, neuronal hypersensitivity underlying sub-acute cough and neuronal hypersensitivity underlying acute cough.
 5. The method of claim 1, wherein about 50 mg of the compound of Formula (I) is administered multiple times daily.
 6. The method of claim 1, wherein about 100 mg of the compound of Formula (I) is administered multiple times daily.
 7. The method of claim 1, wherein about 100-900 mg of the compound of Formula (I) is administered multiple times daily.
 8. The method of claim 1, wherein the cough is reduced by about 75%.
 9. The method of claim 1, wherein R¹ is hydrogen or methyl.
 10. The method of claim 1, wherein R² is aminosulfonyl, haloalkyl, alkylsulfonyl, carboxyalkyl or alkylcarbonyl.
 11. The method of claim 1, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 12. The method of claim 1, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 13. The method of claim 1, wherein the compound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 14. A method for treating a cough in a patient, wherein the cough is selected from the group consisting of chronic pathological cough, sub-acute pathological cough, acute pathological cough, neuronal hypersensitivity underlying chronic cough, neuronal hypersensitivity underlying sub-acute cough and neuronal hypersensitivity underlying acute cough, the method comprising administering to the patient in need thereof an effective amount of a compound having the formula:

or a pharmaceutically acceptable salt thereof.
 15. A method for treating a cough in a patient, wherein the cough is selected from the group consisting of chronic pathological cough, sub-acute pathological cough, acute pathological cough, neuronal hypersensitivity underlying chronic cough, neuronal hypersensitivity underlying sub-acute cough and neuronal hypersensitivity underlying acute cough, the method comprising administering to the patient in need thereof an effective amount of a pharmaceutically acceptable salt of a compound having the formula: 